转基因甘蔗BtG-2的T-DNA侧翼序列分析及其转化事件特异性检测

转基因甘蔗BtG-2是利用农杆菌介导法把Cry1Ac-2A-gna融合抗虫基因导入‘新台糖22号’的转基因甘蔗株系,具有良好的抗虫特性和农艺性状。为了明确转基因甘蔗BtG-2的分子特征及其检测方法,推进其生物安全性评价工作,以BtG-2的T2代为研究材料,利用Southern杂交检测外源基因在转基因甘蔗基因组内的拷贝数;利用染色体步移技术分离外源基因在甘蔗基因组中插入位点的侧翼序列,并建立了该转化体高效灵敏的特异性PCR检测方法。结果表明：Southern杂交检测证明外源T-DNA以单拷贝方式插入BtG-2株系;经过3次的热不对称巢式PCR扩增,获得外源基因T-DNA左边侧翼序列984 bp和右边侧翼序列705 bp;以这2个序列和相应的T-DNA的左右端序列分别设计3对检测引物对,建立了BtG-2株系的转化事件特异性PCR检测方法,扩增效率最高的引物对LS011/LA451和RS160/RA588分别扩增到440 bp和428 bp的特异片段。其中T-DNA左侧设计的LS011/LA451检测引物对扩增的灵敏度高、特异性强,能够在甘蔗BtG-2基因组DNA相对含量为0.1%的模板中检测出转基因目的成分,相当于9个单倍体基因组拷贝数。本研究完成了转基因株系BtG-2的分子特征及其转化事件特异性检测,为该转基因甘蔗及其衍生产品的检测和身份识别提供技术依据。     

Development of colchicine‐induced tetraploid St. Augustinegrass (Stenotaphrum secundatum) lines

St. Augustinegrass is well suited for lawns and commercial landscapes. While many genotypes are cross‐fertile, all cultivars are propagated vegetatively in sod production. To ensure varietal purity, development of sterile triploid hybrids by crossing tetraploid and diploid genotypes has been successfully used in other warm‐season turfgrasses. Applying this model in St. Augustinegrass would be beneficial to sod producers and turf managers who require purity for certification and uniformity for performance, respectively. This study was conducted to develop colchicine‐induced tetraploid lines of St. Augustinegrass. Seeds of cultivar ‘Raleigh’ were treated with four colchicine concentrations at four exposure times. A non‐treated control was included among the treatments. Seedlings that germinated were screened for genome size changes using flow cytometry. Line DSA 13005 and two progeny lines derived through selfing, DSA 16001 and DSA 16016, were corroborated as tetraploids (2n = 4x = 36) through chromosome counts. These lines will be used in future breeding efforts to attempt development of sterile triploid cultivars of St. Augustinegrass.     

Revisiting rice breeding methods – evaluating the use of rapid generation advance (RGA) for routine rice breeding

Rice production needs to increase in the future in order to meet increasing demands. The development of new improved and higher yielding varieties more quickly will be needed to meet this demand. However, most rice breeding programmes in the world have not changed in several decades. In this article, we revisit the evidence in favour of using rapid generation advance (RGA) as a routine breeding method. We describe preliminary activities at the International Rice Research Institute (IRRI) to re-establish RGA on a large scale as the main breeding method for irrigated rice breeding. We also describe experiences from the early adoption at the Bangladesh Rice Research Institute. Evaluation of RGA breeding lines at IRRI for yield, flowering time and plant height indicated transgressive segregation for all traits. Some RGA lines were also higher yielding than the check varieties. The cost advantages of using RGA compared to the pedigree method were also empirically determined by performing an economic analysis. This indicated that RGA is several times more cost effective and advantages will be realized after 1 year even if facilities need to be built. Based on our experience, and previous independent research empirically testing the RGA method in rice, we recommend that this method should be implemented for routine rice breeding in order to improve breeding efficiency.     

Construction and evaluation of introgression lines and fine mapping of ehd8 from Jinghong common wild rice (Oryza rufipogon)

Heading date is one of the most important traits in rice and regulated by multiple genes. Common wild rice is the ancestor of Asian cultivated rice and harbours abundant genetic diversity. To use wild rice resource in rice breeding, a set of 154 introgression lines (ILs) covering 93% of the genome of Jinghong common wild rice was constructed in the background 'Teqing', using 208 simple sequence repeat markers evenly distributed on 12 chromosomes. Among the ILs, the line JIL64 displayed late heading independent of photoperiod. Genetic analysis using the two F₂ populations crossed ''Teqing'/JIL64 and JIL64/'Teqing' revealed that late flowering was controlled by a recessive gene on chromosome 8 (designated early heading date 8, ehd8), and ehd8 was fine mapped to the 50‐kb region flanked by markers RM22221 and 64Indel4. Sequencing and qRT‐PCR demonstrated that LOC_Os08g01410 and LOC_Os08g01420 were deleted in JIL64 and may be associated with the late heading of Jinghong common wild rice. These findings lay a practical foundation for characterizing ehd8, and the ILs help to mine genes from Jinghong common wild rice.     

SDS-PAGE和PCR检测1B/1R易位系研究初报

采用18个冬小麦品种作为供试材料,分别通过SDS-PAGE检测黑麦Secalin蛋白和PCR检测黑麦1RS染色体或其片段,结果表明:SDS-PAGE检测不出不带有secalin蛋白的1B/1R小片段易位系,而PCR则能检测出,从而提高1B/1R易位系的检出率;SDS-PAGE和PCR结合是检测1B/1R易位系的一种快速经济的有效方法。     

Investigations on the Occurrence of Hereditary Diseases in the Danish Cattle Population 1989-1991

Agerholm, J.S., A. Basse and K. Christensen: Investigations on the occurrence of hereditary diseases in the Danish cattle population 1989-1991. Acta vet. scand. 1993, 34, 245-253.– The methods of the Danish Bovine Genetic Disease Programme are outlined, and the results obtained during the first 3 years in function are described. The most common disease reported was spinal muscular atrophy in calves of the Red Danish Dairy breed with 312 reports. Necropsy was performed on 162 cases, and spinal muscular atrophy was diagnosed in 82 of these. Bovine progressive degenerative mye-loencephalopathy, rectovaginal constriction, syndrome of arthrogryposis and palatoschisis, hereditary chondrodysplasia (2 different types), syndactylism, epitheliogenesis imperfecta, and osteogenesis imperfecta was diagnosed with 1 case each. Lethal trait A46 was diagnosed in 4 calves. Some of these diseases have not previously been described in Denmark, and epitheliogenesis imperfecta was for the first time diagnosed in the Hereford breed. Chromosome translocation 1/29 was detected in the Blonde d’Aquitaine (BAQ), Limousine, and Red Danish Dairy breed. The aberration occurred frequently in BAQ. Furthermore, a complex chromosome translocation t(l;8;9)(q45;ql3;q26) was detected in the Red Danish Dairy breed.     

可持续发展类型与测度的理论探讨

可持续发展的类型划分与可持续发展的测度是可持续发展理论研究的重要内容。科学反应可持续发展的特征 ,有利于可持续发展类型划分 ,是可持续发展测度的依据和目标。本文运用人地关系协调分析法与经济分析法 ,特别是与替代弹性不变生产函数分析法 (C.E.S生产函数 )相结合 ,提出了可持续的人均国内生产总值 ,探讨了可持续发展中人文资本、自然资本相互协调、相互依存的关系特征 ,建立可持续发展测度的指标系统和可持续发展的类型体系     

Development of Chinese Near-isogenic Line of Rice and Their Differentiating Ability of Pathogenic Races of Blast Fungus

The rice( Oryza L. ) variety, Lijiangxintuanheigu(LTH), is a universally susceptible variety. A set of near-isogenic lines (NILs) of rice which has single resistance gene for each line was developed by use of LTH as recurrent parent and Kusabue,Tsuyuake, K1, PiNo. 4, K60 and BL1, which are from Kiyosawa‘s differential varieties(KDV), as donor parents. Differentiating ability of these NILs to Philippine isolates was compared with that of their donor parents, KDV and IRRI‘ s NILs. The results of comparative studies were as follows: (1) Differentiating ability of Chinese NILs to Philippine isolates was much stronger than that of their donor parents and that of KDV. (2) Chinese NILs had similar ability to differentiate races to IRRI‘s NILs or higher differentiating ability than them. But IRRI‘s NILs almost had no differentiating ability to isolates from japonica rice-growing regions. They can not be used to differentiate races of isolates from japonica ricegrowing regions. Whereas, Chinese NILs can be used to differentiate races of isolates from both japonica ricegrowing regions and indica rice-growing regions. It was concluded that the set of Chinese NILs can be used as international differentials.     

In vitro Plant Regeneration from the Mature Tissue of Navel Orange (Citrus sinensis L. Osbeck) by Direct Organogenesis

An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments of Newhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growth regulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation is beneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formation efficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAP and 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds per internodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and were transferred to the soil.     

无特定病原体京白鸡的核型研究

采用腹腔注射秋水仙素,取骨髓细胞制备染色体标本,对无特定病原体京白鸡的核型进行了分析.并根据前10对染色体相对长度、臂比值及着丝点指数的测量结果,与已公布的其它地方鸡品种进行了比较.